Improved Escherichia coli Bactofection and Cytotoxicity by Heterologous Expression of Bacteriophage ΦX174 Lysis Gene E

Abstract

Bactofection offers a gene delivery option particularly useful in the context of immune modulation. The bacterial host naturally attracts recognition and cellular uptake by antigen presenting cells (APCs) as the initial step in triggering an immune response. Moreover, depending on the bacterial vector, molecular biology tools are available to influence and/or overcome additional steps and barriers to effective antigen presentation. In this work, molecular engineering was applied using Escherichia coli as a bactofection vector. In particular, the bacteriophage ΦX174 lysis E (LyE) gene was designed for variable expression across strains containing different levels of lysteriolysin O (LLO). The objective was to generate a bacterial vector with improved attenuation and delivery characteristics. The resulting strains exhibited enhanced gene and protein release and inducible cellular death. In addition, the new vectors demonstrated improved gene delivery and cytotoxicity profiles to RAW264.7 macrophage APCs.

In situ pneumococcal vaccine production and delivery through a hybrid biological-biomaterial vector

“…However, the design of the hybrid vector allows both the chemical and biological aspects of the device to extend dosing level safety. Namely, the vector’s polymer coating and the inclusion of a lysis E (LyE) gene provide alternative mechanisms of attenuation ( 3 , 10 ). In the case of LyE, the membrane perforative activity of this protein affords safety of up to 10 20 dose levels for the resulting hybrid device (fig.…”

Bacterial Ghosts-Based Vaccine and Drug Delivery Systems

“…This increases internal osmotic pressures, inducing the release of cell contents [21]. Currently, the most commonly used method for the preparation of BGs involves cloning lysis gene E (276 bp, accession number: MF426914.1) into expression regulation systems of Gram-negative bacteria, which utilizes the switch function of the control element to control expression [22]. The preparation process for BGs is shown in Figure 2.…”

Influence of molecular weight upon mannosylated bio-synthetic hybrids for targeted antigen presenting cell gene delivery

“…Specifically, polymer molecules are believed to first adsorb to the surface of bacteria prior to mediating mild disruptions of the outer and inner Gram-negative leaflet membranes through diffusive mechanisms. Thus, bacterial membrane destabilizations has been associated with improved gene delivery outcomes by facilitating the controlled release of internal protein and DNA cargo [7, 18]. …”